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Alw21I (BsiHKAI)

5'...G W G C W↓C...3'
3'...C↑W C G W G...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Optimal incubation at 37°C O buffer for 100% activity Thermal inactivation at 65°C in 20 min LO certified

The Alw21I (BsiHKAI) restriction enzyme recognizes GWGCW^C sites and cuts best at 37°C in O buffer (Isoschizomers: Bbv12I, BsiHKAI)
  
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Lambda DNA digested with Alw21I (BsiHKAI)

Lambda DNA digested with Alw21I (BsiHKAI), 1% agarose, 28 cleavage sites

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible Ends
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferAlw21I is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue
)1X		 G (green
)1X		 O (orange)
1X		 R (red)
1X		 Tango (yellow)
1X / 2X
O (Orange) 37°C 0-20 20-50 100 50-100 20-50 50-100 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...m5C GWGCWm5C G...3'
3'... Gm5CWCGW Gm5C...5'		 No effect
EcoKI (AAC(N)6GTGC) 5'...Am6AC(N)6G TGCWC...3'
3'...T TG(N)6Cm6ACGWG...5'		 No effect

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
28 3 3 8 5 5
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
4 4 4 4 2 5

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
GAGCA^C
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GAGCA^C)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
GAGCT^C
  • Eco24I (BanII)* (GAGCT^C)
  • SacI/FastDigest SacI (GAGCT^C)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GAGCT^C)
  • AluI/FastDigest AluI
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • CviJI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II
  • Eco24I (BanII)
  • SacI/FastDigest SacI
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • SetI
  • SetI* (AGCT)
  • AluI/FastDigest AluI
  • CviJI
  • SetI
GTGCA^C
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)* (GTGCA^C)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GTGCA^C)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest ApaLI (Alw44I)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I) (ATGCA^T)
  • HpyCH4V
  • PstI/FastDigest PstI (CTGCA^G)
  • SdaI (SbfI)/FastDigest SbfI (SdaI) (CCTGCA^GG)
  • BsgI
  • HpyCH4V
GTGCT^C
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)* (GTGCT^C)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)